From Enzyme Discovery to Engineered Strains: Biocatalytic Solutions for Pharmaceutical Synthesis

Biocatalysis is increasingly adopted in the chemical industry due to its sustainability advantages, such as

biodegradable catalysts, high selectivity, and operation under mild conditions.[1] In particular, the precise control

enzymes exert over reaction outcomes makes them ideally suited for the synthesis of active pharmaceutical

ingredients. In this talk, I will present our integrated approach to developing new enzymatic and microbial tools for the

synthesis of pharmaceutical building blocks.

We focus on the discovery, characterization, and engineering of nitrogen–nitrogen bond-forming enzymes

(NNzymes), which utilize various metallo-cofactors and reaction types to catalyze N–N couplings, forming unique

bioactive molecules.[1] A particular focus lies on piperazate synthases (PZSs) involved in the biosynthesis of rare

nitrogen-containing heterocycles.[2] These enzymes offer a biocatalytic route to L-piperazate—a pharmacophore found

in several bioactive compounds—highlighting the potential of pathway-inspired enzyme engineering. We expand the

scope of characterized PZSs through genome mining and reveal promiscuous activities leading to functionalized

hydrazine derivatives[2] and other nitrogen heterocycles of pharmaceutical relevance.

[3]

Additionally, we explore Rieske non-heme iron oxygenases (ROs)—an underexploited enzyme family known for

their regio- and stereoselective C–H functionalization capabilities. To harness their potential, we have optimized both

in vivo[4] and in vitro[5] RO systems. We are also exploring hybrid RO systems[6] and redox partner fusion proteins[7] to

enhance catalytic activity, opening new avenues for late-stage functionalization in drug synthesis.

Complementing these enzyme-focused efforts, we have developed synthetic biology tools for Cupriavidus

necator,

[8–10] a versatile and robust chemolithoautotrophic bacterium with high potential for C1-based

biomanufacturing. By improving its genetic tractability, we demonstrate how C. necator can be transformed into an

efficient platform for whole-cell biocatalysis in pharmaceutical applications.[11]

Together, these efforts underscore how combining enzyme discovery and engineering with strain development

of non-model bacteria can significantly expand the biocatalytic toolbox for more sustainable and efficient

pharmaceutical synthesis.